Use the following information to draw and label a gel assume...

Use the following information to draw and label a gel. Assume that the electrophoresis step is completed and that you are viewing the gel on the UV transilluminator. • Indicate where the cathode and anode were in relationship to the gel. • A 100 bp DNA ladder was added to lane 1. This DNA ladder contains 10 different sizes of DNA fragments in 100 bp intervals from 100 bp to 1000 bp. Draw these 10 bands in lane 1, and label the size of each band. • The PCR product from a reaction to make one 350 BP Fragment was added to lane 2. Draw a band in lane 2 representing a successful PCR reaction, and label the size of the band. For lanes 3-5 an experiment was run. There was a linear DNA fragment that was 900 BP. Lane 3: The fragment was not digested Lane 4: The fragment was digested with a RE (RE#1) cutting at the exact midpoint of the fragment. Lane 5: A double digest (RE#1 and RE#2) was completed and the following fragments show up on the gel : 450, 300 and 150. Roughly map the RE sites on this fragment (note there are two possible answers, include both).

Transcript

00:01 The question asks you to use following information to drill and label a gel.

00:05 So assuming that the electrophoresate step is completed and you are viewing the gel on a uv truss illuminator, indicate where the cathode and an anode when were, first of all, in relation to the gel.

00:21 So dna always run from negative to positive.

00:25 So i have already labeled that.

00:28 And the first lane is dna letter added to lane one.

00:35 So you have a total of lane five.

00:37 So i'm going to write down one, two, three, four, and five.

00:44 So lane one is the dna letter.

00:49 Contain 10 different sizes of dna fragment in 100 base pair intervals from 100 base pair to 1 ,000 base pair.

00:55 So draw these 10 bands in lane 1.

00:58 And label the size of each band.

01:01 So basically in lane one you have a dna ladder.

01:07 The longest bend is going to be on top of the gel because the dna run for negative to positive.

01:13 The longest fragment is going to run closer to the well.

01:19 One, two, three, four, five, six, seven, eight, nine.

01:35 So the top one is 1 ,000 base pair.

01:42 900, 800, 700.

01:50 You can see it goes from the longest to the shortest.

01:55 The next one, 600, 500, 400, 300, 300, 300, 200, and the shortest is 100, which is at the bottom of the job because it runs the fastest.

02:13 Okay, so now that's lane one.

02:17 The pcr product from a reaction to make a 350 base pair fragment was added to lane 2.

02:24 So lane 2 is the pcr product, which will have a size of 350 base pair fragment.

02:31 So draw the band in lane 2, present a successful pcr reaction and label the size of the band.

02:37 Now, this band is 350, so that means it's going to run at the position between 3 .3 .5.

02:46 And 400, so it's in between somewhere here.

02:51 Alright, so now let me try to measure around here.

03:04 So this band here is about 350 base pair.

03:12 It somehow runs between 300, 400 base pair bend.

03:17 Okay, so now, from lay 3 to 5, our experimental was run.

03:22 There was a linear dna fragment that was a total of 900 base pair.

03:26 Lane 3, the fragment was not digested.

03:30 So that means this ban is going to be right at the 900 base pair position.

03:39 So somewhere here, it should line up with exactly the 900 base pair band compared to lane 1.

03:47 So 3 is undigested.

03:50 Then lane 4 is a fragment that was digested.

03:58 With restriction number one.

04:01 This is undigested.

04:04 This is restriction one.

04:09 Cutting the exact midpoint of the fragment...

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